Evaluation of the protective effect of vitamin C on

hepatic damage caused by formaldehyde in rats

Hasan Amiri

, Niloofar Ghodrati

, Amir Noyani

*, Saeed Abbasi

, Hamed

Basir Ghafouri

and Bahareh Seyyed Salehi

MD, Emergency Medicine Management Research Center, Iran University of Medical Sciences, Tehran, Iran

MD, Alborz University of Medical Sciences, Tehran, Iran

MD, Trauma and Injury Research Center, Iran University of Medical Sciences, Tehran, Iran

ABSTRACT

Formaldehyde (methanol) is a chemical substance, with nasty smelling which is normally used to  x cadavers and histological

process. The main use of formaldehyde is in production of synthetic resins, wood-plastic products and  ber industry. Formalde-

hyde has a negative impact on performance of organ system with oxidative stress in the body .specially on the hepatic. Some stud-

ies have demonstrated that antioxidants such as vitamin C can play an important role to protect from damage caused by harmful

substance. The aim of this study was to evaluate the protective effect of vitamin C on hepatic damage caused by formaldehyde in

rats.This study included 24 adult male rats with weighing 250-300 gr. rats were randomly divided into 3 groups each comprising 8

rats. 1- Control group: exposed to normal saline (1cc/kg per day for 10 days) that injected intraperitoneally, 2 -groups E1: exposed

to formaldehyde 10mg/kg per day for 10 days, 3 - group E2: simultaneously exposed to vitamin C (200mg/kg) per day for 10 days&

formaldehyde 10 mg/kg intraperitoneally 10 days. At the end of the exposure period, the mice were placed under anesthesia and

surgery. The blood samples were taken for isolation of serum and measurement of AST and ALT in.In this study, AST and ALT

levels of 24 adult rats in the all groups were measured and the results were compared. The amount of AST in the all groups was

signi cant (P <0.05). The amount of AST in group E2 compared to controls was signi cant (P = 0<0.05) and the amount of AST in

group E2 compared to E1 was not signi cant (P = P>0.05). Also, comparison of the results showed signi cant difference in serum

ALT in third group. Serum ALT level in E1 compared to control group was signi cant (P <0.05) and ALT in group E2 compared to

control group was signi cant (P <0.05). The amount of ALT in group E2 compared to E1 was not signi cant (P = 0.847).According

to this study, intraperitoneal administration of formaldehyde at a dose of 10 mgr/kg per 10-day can increase AST and ALT and

vitamin C at a dose of 200 (mg / kg) per day for 10-days can reduce amount of AST and ALT in adult rats exposed to formaldehyde

KEY WORDS: FORMALDEHYDE, AST, ALT, LIVER

213

ARTICLE INFORMATION:

*Corresponding Author: amir_noiany@yahoo.com

Received 27

Dec, 2016

Accepted after revision 2

March, 2017

BBRC Print ISSN: 0974-6455

Online ISSN: 2321-4007

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Online Contents Available at: http//www.bbrc.in/

Biosci. Biotech. Res. Comm. Special Issue No 1:213-217 (2017)

214 EVALUATION OF THE PROTECTIVE EFFECT OF VITAMIN C ON HEPATIC DAMAGE BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

Hasan Amiri et al.

INTRODUCTION

Formaldehyde is a chemical material with a nasty smell,

which is used in  xing cadavers, histological processes,

synthetic resins, wooden and plastic productions and

industrial  ber production. It has a negative effect on

performance of body organs. It is metabolized to formic

acid by dehydrogenase formaldehyde and dehydrogenase

aldehyde enzymes in the liver and erythrocyte which is

exorcised through urine, excretion and breathing with

different macromolecules such as protein, acid nucleic or

interact with the light molecules like amino acid. Formal-

dehyde can cause oxidative stress in the body and has

a bad effect on respiratory system and blood circulation

and liver (Golalipour et al., 2007 Mendis et al., 2007,

Cheng et al., 2003, Collins and Lineker 2004 Kini et al.,

2004). Formaldehyde was known as a harmful factor for

liver cells in the different studies. Antioxidants are such

components that help body to destroy free radicals oxi-

dative stress is actually imbalance between oxidants and

antioxidants. When the amount of oxidants increases,

the cells are damaged. Antioxidants include Vitamin

A, E, C, Zn and selenium, which play a crucial role on

inhibition of free radicals and stability of cell membrane,

(Kini et al., 2001, Gupta et al., 2004, Woolaqrd et a;

2002).

Vitamin C (Ascorbic Acid) is a white or yellow odor-

less solid substrate with the molecular formulation of

C6H806. Ascorbic Acid is produced in the liver of plants

and animals (except some special kind and human). Vita-

min C operates as antioxidants in the body and cause

acceleration of Fe, Cu and also revives of folic acid and

collagen making. Vitamin C as a soluble antioxidant

becomes active by moving oxygen and nitrogen ele-

ments. The role of vitamin E as a protective agent on

liver damage caused by formaldehyde in the previous

studies was approved. The role of A,E,C Vitamins as

protective antioxidants on liver hepatotoxicity was also

investigated. It is possible to evaluate liver function by

checking the indicators in the blood, (Kum et al., 2011,

Kini et al., m 2011, Djeffal et al., 2011, Ememghorashy

et al., 2012).

With regard to the formaldehyde effects in making

oxidative stress, free radicals and their effect on liver

performance and in accordance with the performed

studies related to the formaldehyde on the liver tis-

sue changes as glomerular vascular congestion and

also minor decline holes in the pipe cells and stud-

ies about the protection effect of Vitamin C on pre-

vention of formaldehyde damage, in this study, we

decided to investigate the protection effect of Vita-

min C on liver performance of rats after exposing to

formaldehyde.

METHODOLOGY

In this study 24 adult rat of Wistar bread were selected

and divided into 3 equal groups. The  rst group (c)

received 1cc/kg of normal saline and the second group

(E2) received 10 mg/kg formaldehyde and the third group

(E3) received both 10mg/kg formaldehyde and 200 mg/

kg Vitamin C in 10 days by injection into peritoneal.

During the study, the rats were under normal condition

of shelter with 24±2 temperature and appropriate feed-

ing. 3 weeks after  nishing the injection phlebotomy,

after anesthesia by sterile syringe was performed. After

making  occulation by centrifuge, samples were sepa-

rated with 1500 rpm during 10 minutes and they were

kept in -20 c till testing for evaluation of ALT and AST.

The amount of ALT and AST were measured by using

Pars Azmoon kits and auto analyzer tool (BiotecticaIn-

strument BT 1000).

Data analysis was performed after entering the data

to SPSS Ver.20program and to investigate normality of

cantinas quantitative data distribution, Shapiro-wilk

test was conducted. The explicit result of the groups was

reported as average and deviation from standard devia-

tion. The comparison between the investigated groups

was performed by using ANOVA test. If the average dif-

ferences of statistic results were meaningful the LSD test

were used to compare them. The meaningful level was

considered less than 0.05 (P<0.05).

RESULTS AND DISCUSSION

In this study, 24 adult male rats of Wistar bread were

involved. Among them 8 rats (33%) considered as con-

trol group, 8 rats (33%) considered as test group 2 (E2)

and another 8 rats (33.3%) as test group 2 (E2). Descrip-

tive factors and the comparison between the weights of

rats among 3 groups of control E1, E2 was shown in the

table 1. In order to compare AST level in 3 groups of

control (c) E1,E2. Variance analysis test (ANOVA) was

conducted and in order to compare 2 group’s level of

ALT, the LSD test was came out. Descriptive speci ca-

tions and the comparison between ALT levels of three

groups include group c, E1, E2 were listed in the table 2.

According to the comparison of AST level, it was sug-

gested that there is a signi cant difference in 3 groups

of control, E1, E2 (p<0.05). Also, with regard to the 2

groups comparison, The difference between the AST

level of group c and group E1 was meaning (p<0.05)

and the difference between AST level of control (c) and

test group (E1) was also meaningful (p<0.05). However,

there is not a meaningful difference between AST level

of study group E1 and E2 (p>0.05) (plot 1). In order to

compare, ALT level in 3 study groups of control (c), E1,

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS EVALUATION OF THE PROTECTIVE EFFECT OF VITAMIN C ON HEPATIC DAMAGE 215

Hasan Amiri et al.

PLOT 1. AST level average plot for 3 study groups of control

(c), Test 1 (E1), Test2 (E2)

PLOT 2. ALT level average plot of 3 study groups of control

(c), Test 1 (E1), Test 2 (E2)

Table 1. Rats weight in 3 study groups of control (c), E1, E2

Speci cation

P-Value

Control Test 1 Test 2

Weight (gr)

Standard deviation ± Mean

Range

214,37 ± 11,01

202-235

212,12±2,79

208-217

215,5±3,96

211-222

0.62

Table 2. AST amount of 3 study groups of control (c), Test 1 (E1), Test 2 (E2)

speci cation Group Comparison among groups

Control E1 E2 Statistic F P-Value

AST

Mean±SD

Range

78.5±5.75

68-86

126.5±18.08

96-145

110.75±14.81

86-136

24.78 <0.001

E2. The one way variance analysis (ANOVA) was con-

ducted and in order to compare ALT level between 2

groups, the LSD test was used.

Descriptive speci cations and the comparison of ALT

level among 3 study groups of control (c), test 1 (E1) and

test 2 (E2) was shown in the table 3. According to the

comparison of three groups, the ALT level has a mean-

ingful difference (p<0.05). It was also suggested that

there is a signi cant difference between ALT level of

control group (c) and test 1 group (E1). (p<0.05) and ALT

Hasan Amiri et al.

216 EVALUATION OF THE PROTECTIVE EFFECT OF VITAMIN C ON HEPATIC DAMAGE BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

level of C and E2 group (p<0.05) and also between E1,E2

groups (p<0.05).

Formaldehyde can affect the body organs due to cre-

ating free radicals. Formaldehyde can also affect the

kidney performance such as excretion of waste sub-

stances. Antioxidant can prevent the harmful effect of

formaldehyde . Vitamins are one of the most important

antioxidants – especially Vitamin C – which play an

effective role in protection against oxidative damage

caused by free radicals . On the other hand, this Vitamin

can enhance the activity of antioxidant enzymes in the

liver tissue,( Claudia et al., 2003, HaiXia et al., 2012,

Ukmali and Armutcuf 2011, Sajadi et al., 2008, Shang

et al., 2014).

18-WILSON ET AL., 2014)

According to the above description, 24 rats of Wistar

bread were involved in this study with regard to the pre-

vious studies and the ability and facility of laboratory

and human resource present in the research center. This

study was similar to the study of Ukmali et al. (2011).

Histology of liver cells was just investigated and no

treatment was suggested to prevent formaldehyde effect.

In the study of Ukmali some evidences were shown

about in ammation in the liver cells .

On the other hand this study was similar to that of

Shang et al (2014), except that in this study, Vitamin C

was fed to the rats and histology of kidney and liver cells

was investigated. With regard to the performed studies

and the side effect of formaldehyde which was approved

by them and also concerning this fact that people and

doctors are always exposed to this substrate without

being able to perform histology investigations on human

body, the liver enzymes was measured to evaluate ALT

and AST indicators. It was shown that there were a sig-

ni cant difference between the control groups and the

groups exposed to formaldehyde which can be related to

the in ammatory response of liver cells to formaldehyde

injection. Standardization was performed according to

the weight, gender and bread. So, the meaningful differ-

ence statistically approved the harmful effect of expos-

ing to this substrate (p<0.0001). On the other hand, ALT

and AST level in the E2 group treated by vitamin C were

decreased but not to the normal level of liver enzymes

which is probably because of high toxicity effect of for-

maldehyde. More reduction may be reached by increas-

ing the time of treatment or Vitamin C dose.

CONCLUSION

According to this study, intraperitoneal administration

of formaldehyde at a dose of 10 mge/kg per 10-day can

increase AST and ALT and vitamin C at a dose of 200

(mg / kg) per day for 10-days can reduce amount of AST

and ALT in adult rats exposed to formaldehyde.

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